RESUMO
Ultradeformable liposomes (UDLs) containing sodium cholate as edge activator could be an appropriate skin drug-delivery system for chloroaluminum phthalocyanine (ClAlPc) during photodynamic therapy (PDT) against cutaneous leishmaniasis (CL). The aim of this work was to study cell internalization, reactive oxygen species (ROS) production, and toxicity/genotoxicity and transdermal delivery of UDL-ClAlPc, and to determine whether PDT was able to induce anti-leishmanial activity in Leishmania (Viannia) braziliensis experimental models. Prepared liposomes had an average size of 118.39â¯nm, zeta potential of -37.83â¯mV, and polydispersity index of 0.15. Liposomal internalization (red fluorescence inside cells), ROS generation (green fluorescence by 2,7-dichlorodihydrofluorescein diacetate [DCFH-DA] cleavage) and non-specific DNA damage (photo-comets) were observed after PDT. Transdermal delivery of ClAlPc, measured by in vitro diffusion experiments through BALB/c skin, showed that UDL-ClAlPc was able to deliver very low quantities of ClAlPc (<1%) to deep skin layers. PDT using UDL-ClAlPc induced photodamage in mammalian cells (J774, THP-1, and NIH-3T3), promastigotes, and intracellular amastigotes without a selective response against amastigotes (selective index ≥1). Topical once-daily ClAlPc-UDL plus visible-light irradiation (20â¯J/cm2) twice weekly for 3 weeks was ineffective against L. (V.) braziliensis-infected BALB/c mice, whereas miltefosine 30â¯mg/kg/day orally for 10 days healed the lesions and scars, without parasites observed on the slides. Even though UDLs preserved ClAlPc photoactivities and were able to deliver ClAlPc to dermis, they were unable to result in healing of CL-infected mice after PDT. Experiments using different CL animal models and liposomes with increased skin permeability abilities are recommended.
Assuntos
Indóis/administração & dosagem , Leishmania braziliensis/efeitos dos fármacos , Leishmaniose Cutânea/tratamento farmacológico , Compostos Organometálicos/administração & dosagem , Fotoquimioterapia/métodos , Animais , Linhagem Celular , Dano ao DNA , Feminino , Concentração Inibidora 50 , Lipossomos , Camundongos , Camundongos Endogâmicos BALB C , Espécies Reativas de Oxigênio/metabolismoRESUMO
The effectiveness of various pharmaceutical formulations of ketoconazole was evaluated in experimental models of cutaneous leishmaniasis (LC) in BALB C mice. Topical gel, lipogel, and cream formulations containing permeation enhancers and different concentrations of ketoconazole were prepared. Stability, toxicity and anti-Leishmania activity were determined in vitro. In addition, the effectiveness of topically applied formulations in LC-infected mice infected with Leishmania (Viannia) braziliensis was evaluated in vivo. Cream formulations were additionally evaluated in mice infected with L. (V.) panamensis. The systems evaluated maintained in vitro the activity of ketoconazole against parasites; however, none of the formulations were effective in curing LC lesions in mice. Topical treatment with miltefosine (used as a control) cured the lesions. It is concluded that the ketoconazole-containing formulations designed in this study were not effective against LC in infected mice.
Se evaluó la efectividad de diversas formulaciones farmacéuticas de ketoconazol en modelos experimentales de leishmaniasis cutánea (LC) en ratones BALB C. Fueron preparadas formulaciones tópicas tipo gel, lipogel y crema conteniendo potenciadores de la permeación y diferentes concentraciones de ketoconazol. Se determinó la estabilidad, la toxicidad y la actividad anti-Leishmania in vitro. Además, se evaluó in vivo la efectividad de las formulaciones aplicadas tópicamente en ratones con LC infectados con Leishmania (Viannia) braziliensis. Las formulaciones tipo crema fueron evaluadas adicionalmente en ratones infectados con L. (V.) panamensis. Los sistemas evaluados mantuvieron in vitro la actividad del ketoconazol contra los parásitos; sin embargo, ninguna de las formulaciones fue efectiva en curar las lesiones de LC en los ratones. El tratamiento tópico con miltefosina (utilizado como control) curó las lesiones. Se concluye que las formulaciones que contienen ketoconazol diseñados en este estudio, no fueron efectivos contra la LC en los ratones infectados.
Assuntos
Cetoconazol/administração & dosagem , Leishmaniose Cutânea/tratamento farmacológico , Administração Tópica , Animais , Modelos Animais de Doenças , Composição de Medicamentos , Humanos , Camundongos Endogâmicos BALB C , Resultado do Tratamento , Adulto JovemRESUMO
Resumen Introducción: La diversidad de las formas clínicas de la leishmaniasis del Nuevo Mundo (desde formas cutáneas localizadas a diseminadas o formas mucosas) causada por especies del subgénero Viannia podría inferir en la eficacia de los tratamientos tópicos. El objetivo del presente trabajo fue determinar las características de la leishmaniasis cutánea producida por infecciones con Leishmania (V.) braziliensis y L.(V.) panamensis en ratones BALB/c y la eficacia de un mismo tratamiento tópico. Materiales y métodos: Después de la infección con cada una de las especies se realizó seguimiento de las lesiones determinando su tamaño (mm ) y características macroscópicas, cada siete días por 150 días. Las características histopatológicas (en lesiones y órganos) fueron determinadas 70, 106 y 150 días post-infección y la eficacia de un tratamiento tópico (cura de lesión y parasitológica) fue determinada después del tratamiento con un gel de miltefosina aplicado una vez al día por 20 días sobre las lesiones. Resultados: Se observó un aumento del tamaño de las lesiones en ambos grupos de ratones, sin embargo, un mayor tamaño de las lesiones e intensidad de la respuesta inflamatoria con menos alteraciones epidérmicas fue encontrada en los ratones infectados con L. (V.) braziliensis. En ningún grupo se encontraron parásitos en órganos (nódulos, bazo e hígado) ni diferencias en la efectividad del tratamiento tópico utilizado. Conclusión: La eficacia del tratamiento tópico utilizado no fue afectada por las diferencias macro y microscópicas encontradas en la leishmaniasis producida por las dos especies de Leishmania evaluadas.
Abstract Introduction: The efficacy of topical treatments could be affected by the diversity of clinical forms (localized or disseminated cutaneous forms, mucosal forms) of New World-leishmaniasis caused by species of Leishmania from the subgenus Viannia. The aim of this study was to determine the cutaneous leishmaniasis features produced after infection with Leishmania (V.) braziliensis and L. (V.) panamensis in BALB/c mice and to determine the efficacy of one topical treatment. Materials and methods: Cutaneous leishmaniasis lesions were followed up after infection determining their lesion-size (mm2) and other macroscopic characteristics every 7 days for 150 days. Histopathological patterns (in lesions and organs) were determined 70, 106 and 150 days post-infection and the efficacy (lesion and parasitological cure) of miltefosine gel applied topical once a day for 20 days was determined. Results: An increase of size-lesions was observed in both groups of mice, however, a higher lesion- size and inflammatory response but lower epidermal changes were observed in L. (V.) braziliensis compared with L. (V.) panamensis infected ones. No parasites were observed in organs (nodules, spleen and liver) and no differences were observed in the effectiveness of the used topical treatment. Conclusion: The efficacy of the topical treatment used was not affected by the macro and microscopic differences produced after infection by the two Leishmania species evaluated.
Assuntos
Animais , Leishmania braziliensis , Leishmania guyanensis , Camundongos Endogâmicos BALB C , Anti-Infecciosos LocaisRESUMO
RESUMEN Se evaluó la efectividad de diversas formulaciones farmacéuticas de ketoconazol en modelos experimentales de leishmaniasis cutánea (LC) en ratones BALB C. Fueron preparadas formulaciones tópicas tipo gel, lipogel y crema conteniendo potenciadores de la permeación y diferentes concentraciones de ketoconazol. Se determinó la estabilidad, la toxicidad y la actividad anti-Leishmania in vitro. Además, se evaluó in vivo la efectividad de las formulaciones aplicadas tópicamente en ratones con LC infectados con Leishmania (Viannia) braziliensis. Las formulaciones tipo crema fueron evaluadas adicionalmente en ratones infectados con L. (V.) panamensis. Los sistemas evaluados mantuvieron in vitro la actividad del ketoconazol contra los parásitos; sin embargo, ninguna de las formulaciones fue efectiva en curar las lesiones de LC en los ratones. El tratamiento tópico con miltefosina (utilizado como control) curó las lesiones. Se concluye que las formulaciones que contienen ketoconazol diseñados en este estudio, no fueron efectivos contra la LC en los ratones infectados.
ABSTRACT The effectiveness of various pharmaceutical formulations of ketoconazole was evaluated in experimental models of cutaneous leishmaniasis (LC) in BALB C mice. Topical gel, lipogel, and cream formulations containing permeation enhancers and different concentrations of ketoconazole were prepared. Stability, toxicity and anti-Leishmania activity were determined in vitro. In addition, the effectiveness of topically applied formulations in LC-infected mice infected with Leishmania (Viannia) braziliensis was evaluated in vivo. Cream formulations were additionally evaluated in mice infected with L. (V.) panamensis. The systems evaluated maintained in vitro the activity of ketoconazole against parasites; however, none of the formulations were effective in curing LC lesions in mice. Topical treatment with miltefosine (used as a control) cured the lesions. It is concluded that the ketoconazole-containing formulations designed in this study were not effective against LC in infected mice.